The Western blot technique for detecting and identifying specific proteins in biological samples is commonly used among life scientists. This blotting technique utilizes gel electrophoresis to fractionate native or denatured proteins based on their migration speed (mobility) in a gel while under an electrical field. The proteins, trapped and size-fractionated in a gel, are transferred to and immobilized by a positively-charged membrane. (See FIG. 1) Using primary antibodies specific to the target proteins, as well as secondary antibodies for signal amplification and visualization, expression and modification of the target proteins can be investigated. Although the procedure is well-established, and many tools are available for gel fractionation and blotting, one of the bottlenecks is its limited efficiency and controllability during the blotting procedure.
In most applications of Western blotting, there is a need to evaluate expression levels of multiple protein targets. Signal transduction pathways, for instance, are often activated by protein modifications such as glycosylation and phosphorylation. It is important to be able to determine the amount of signaling molecules that are, in many cases, phosphorylated. However, the current blotting technique can generate only a single membrane per gel, and thus life scientists usually have to prepare multiple gels. When the results from multiple gels are compared, one source of error may arise from potential chemical and physical variations among the gels. Furthermore, since the preparation of protein samples, running gels, and transferring to membranes are time consuming and costly, it is desirable to develop a blotting procedure that enables multiple blotting from a single gel. Such a procedure could reduce potential inconsistencies during sample loading, electrophoresis, and blotting, which are affected by varying factors including operational time, temperature, and chemical composition.
Thus, there is a need for a quicker and more efficient protein separation technique. The present novel technology addresses this need.